This invention relates generally to novel cyanine and indocyanine dyes for use in imaging, diagnosis and therapy. Particularly, this invention relates to compositions of cyanine and indocyanine dyes wherein novel carbocyclic and heterocyclic moieties are incorporated into the polyene portion of the dye molecules.
Several dyes that absorb and emit light in the visible and near-infrared region of electromagnetic spectrum are currently being used for various biomedical applications due to their biocompatibility, high molar absorptivity, or high fluorescence quantum yields. The high sensitivity of the optical modality in conjunction with dyes as contrast agents parallels that of nuclear medicine and permits visualization of organs and tissues without the undesirable effect of ionizing radiation. Cyanine dyes with intense absorption and emission in the near-infrared (NIR) region are particularly useful because biological tissues are optically transparent in this region (B. C. Wilson, Optical properties of tissues. Encyclopedia of Human Biology, 1991, 5, 587-597). For example, indocyanine green, which absorbs and emits in the NIR region has been used for monitoring cardiac output, hepatic functions, and liver blood flow (Y-L. He, H. Tanigami, H. Ueyama, T. Mashimo, and I. Yoshiya, Measurement of blood volume using indocyanine green measured with pulse-spectrometry: Its reproducibility and reliability. Critical Care Medicine, 1998, 26(8), 1446-1451; J. Caesar, S. Shaldon, L. Chiandussi, et al., The use of Indocyanine green in the measurement of hepatic blood flow and as a test of hepatic function. Clin. Sci. 1961, 21, 43-57) and its functionalized derivatives have been used to conjugate biomolecules for diagnostic purposes (R. B. Mujumdar, L. A. Ernst, S. R. Mujumdar, et al., Cyanine dye labeling reagents: Sulfoindocyanine succinimidyl esters. Bioconjugate Chemistry, 1993, 4(2), 105-111; Linda G. Lee and Sam L. Woo. xe2x80x9cN-Heteroaromatic ion and iminium ion substituted cyanine dyes for use as fluorescent labelsxe2x80x9d, U.S. Pat. No. 5,453,505; Eric Hohenschuh, et al. xe2x80x9cLight imaging contrast agentsxe2x80x9d, WO 98/48846; Jonathan Turner, et al. xe2x80x9cOptical diagnostic agents for the diagnosis of neurodegenerative diseases by means of near infra-red radiationxe2x80x9d, WO 98/22146; Kai Licha, et al. xe2x80x9cIn-vivo diagnostic process by near infrared radiationxe2x80x9d, WO 96/17628; Robert A. Snow, et al., Compounds, WO 98/48838).
A major drawback in the use of cyanine dye derivatives is the potential for hepatobilliary toxicity resulting from the rapid clearance of these dyes by the liver (G. R. Cherrick, S. W. Stein, C. M. Leevy, et al., Indocyanine green: Observations on its physical properties, plasma decay, and hepatic extraction. J. Clinical Investigation, 1960, 39, 592-600). This is associated with the tendency of cyanine dyes to form aggregates in solution which could be taken up by Kupffer cells in the liver. Various attempts to obviate this problem have not been very successful. Typically, hydrophilic peptides, polyethyleneglycol or oligosaccharide conjugates have been used but these resulted in long-circulating products which are eventually cleared by the liver. Another major difficulty with current cyanine and indocyanine dye systems is that they offer a limited scope in the ability to induce large changes in the absorption and emission properties of these dyes. Attempts have been made to incorporate various heteroatoms and cyclic moieties into the polyene chain of these dyes (L. Strekowski, M. Lipowska, and G. Patonay, Substitution reactions of a nucleofugal group in hetamethine cyanine dyes. J. Org. Chem., 1992, 57, 4578-4580; N. Narayanan, and G. Patonay, A new method for the synthesis of heptamethine cyanine dyes: Synthesis of new near infrared fluorescent labels. J. Org. Chem., 1995, 60, 2391-2395; E. Fung and R. Rajagopalan, Monocyclic functional dyes for contrast enhancement in optical imaging, U.S. Pat. No. 5,732,104; R. Rajagopalan and E. Fung, Delta1,6 bicyclo[4,4,0] functional dyes for contrast enhancement in optical imaging, U.S. Pat. No. 5,672,333; R. Rajagopalan and E. Fung, Tricyclic functional dyes for contrast enhancement in optical imaging, U.S. Pat. No. 5,709,845) but the resulting dye systems do not show large differences in absorption and emission maxima, especially beyond 830 nm where photacoustic diagnostic applications are very sensitive. They also possess prominent hydrophobic core which enhances liver uptake. Further, most cyanine dyes do not have the capacity to form dendrimers which are useful in biomedical applications.
Therefore, there is a need to design novel dyes that could prevent dye aggregation in solution, predisposed to form dendrimers, capable of absorbing or emitting beyond 800 nm, possess desirable photophysical properties, and endowed with tissue-specific targeting capability.
The publications and other materials used herein to support the background of the invention or provide additional details respecting the practice, are incorporated by reference.
The present invention relates particularly to the novel composition comprising cyanine dyes of general formula 1
wherein a1 and b1 vary from 0 to 5; W1 and X1 may be the same or different and, are selected from the group consisting of xe2x80x94CR10R11, xe2x80x94Oxe2x80x94, xe2x80x94NR12, xe2x80x94Sxe2x80x94, and xe2x80x94Se; Q1 is a single bond or is selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, xe2x80x94Sexe2x80x94, and xe2x80x94NR13; Y1 and Z1 may be the same or different and are selected from the group consisting of xe2x80x94(CH2)cxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)dxe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)exe2x80x94NH2, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94(CH2)fxe2x80x94CH2xe2x80x94NH2, xe2x80x94(CH2)gxe2x80x94N(R14)xe2x80x94(CH2)hxe2x80x94CO2H, and xe2x80x94(CH2)ixe2x80x94N(R15)xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)jxe2x80x94CH2xe2x80x94CO2H; R1 and R10 to R15 may be same or different and are selected from the group consisting of -hydrogen, C1-C10 alkyl, C1-C10 aryl, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, xe2x80x94CH2(CH2xe2x80x94Oxe2x80x94CH2)cxe2x80x94CH2xe2x80x94OH, C1-C20 polyhydroxyalkyl, C1-C10 polyhydroxyaryl, xe2x80x94(CH2)dxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)lxe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)fxe2x80x94NH2, and xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)gxe2x80x94CH2xe2x80x94NH2; c, e, g, h, and i vary from 1 to 10; d, f and j vary from 1 to 100; and R2to R9 may be the same or different and are selected from the group consisting of hydrogen, C1-C10 alkyl, C1-C10 aryl, hydroxyl, C1-C10 polyhydroxyalkyl, C1-C10 alkoxyl, amino, C1-C10 aminoalkyl, cyano, nitro and halogen.
The present invention also relates to the novel composition comprising indocyanine dyes of general formula 2
wherein a2 and b2 are defined in the same manner as a1 and b1; W2 and X2 are defined in the same manner W1 and X1; Q2 is defined in the same manner as Q1; R16 and R10 to R15 are defined in the same manner as R1 and R10 to R15; Y2 is defined in the same manner as Y1; Z2 is defined in the same manner as Z1; and R17 to R28 are defined in the same manner as R2 to R9.
The present invention also relates to the novel composition comprising cyanine dyes of general formula 3
wherein a3 and b3 are defined in the same manner as a1 and b1; W3 and X3 are defined in the same manner W1 and X1; Y3 is defined in the same manner as Y1; Z3 is defined in the same manner as Z1; A1 is a single or a double bond; if A1 is a single bond, then B1 and C1 may the same or different and are selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, xe2x80x94Sexe2x80x94, xe2x80x94Pxe2x80x94, and xe2x80x94NR38 and D1 is selected from the group consisting of xe2x80x94CR39R40, and xe2x80x94Cxe2x95x90O; if A1 is a double bond, then B1 is selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, xe2x80x94Sexe2x80x94, xe2x80x94Pxe2x80x94 and xe2x80x94NR38, C1 is nitrogen or xe2x80x94CR41, and D1 is xe2x80x94CR42; R29 to R37 are selected from the group consisting of hydrogen, C1-C10 alkyl, C1-C10 aryl, hydroxyl, hydrophilic peptide, C1-C10 polyhydroxyalkyl, C1-C10 alkoxyl, cyano, nitro, halogen and xe2x80x94NR43R44; R38 to R42 may be same or different and are selected from the group consisting of-hydrogen, C1-C10 alkyl, C1-C10 aryl, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, xe2x80x94CH2(CH2xe2x80x94Oxe2x80x94CH2)cxe2x80x94CH2xe2x80x94OH, C1-C20 polyhydroxyalkyl, C1-C10 polyhydroxyaryl, xe2x80x94(CH2)dxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)exe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)fxe2x80x94NH2, and xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)gxe2x80x94CH2xe2x80x94NH2; c, e, g, h, and i vary from 1 to 10; d, f and j vary from 1 to 100; R43 and R44 may be the same or different and are selected from the group consisting of hydrogen, C1-C10 alkyl, C1-C10 aryl, or may together form a 5, 6, or 7 membered carbocyclic ring or a 5, 6, or 7 membered heterocyclic ring optionally containing one or more oxygen, nitrogen, or a sulfur atom.
The present invention also relates to the novel composition comprising indocyanine dyes of general formula 4 wherein a4 and b4 are defined in the same manner as a1 and b1; W4 and X4 are defined in the same manner as W1 and X1; Y4 is defined in the same manner as Y1; Z4 is defined in the same manner as Z1; A2 is defined in the same manner as A1; B2, C2, and D2 are defined in the same manner as B1, C1, and D1; and R45 to R57 are defined in the same manner as R29 to R37. 
The present invention also relates to the novel composition comprising cyanine dyes of general formula 5. 
wherein a5 is defined in the same manner as A1; W5 and X5 are defined in the same manner W1 and X1; Y5 is defined in the same manner as Y1; Z5 is defined in the same manner as Z1; A3 is defined in the same manner as A1; B3, C3, and D3 are defined in the same manner as B1, C1, and D1; and R58 to R66 are defined in the same manner as R29 to R37.
The present invention also relates to the novel composition comprising cyanine dyes of general formula 6. 
wherein a6 is defined in the same manner as A1; W6 and X6 are defined in the same manner as W1 and X1; Y6 is defined in the same manner as Y1; Z6 is defined in the same manner as Z1; A4 is defined in the same manner as A1; B4, C4, and D4 are defined in the same manner as B1 , C1, and D1; and R67 to R79 are defined in the same manner as R29 to R37.
This invention is also related to the method of conjugating the dyes of this invention to peptides or biomolecules by solid phase synthesis.
This invention is also related to the method of preventing fluorescence quenching. It is known that cyanine dyes generally form aggregates in aqueous media leading to fluorescence quenching. We observed that in instances where the presence of hydrophobic core in the dyes lead fluorescence quenching, the addition of a biocompatible organic solvent such as 1-50% dimethylsulfoxide (DMSO) restored the fluorescence by preventing aggregation and allowed in vivo organ visualization.
The novel compositions of the present invention comprising dyes of formulas 1 to 6 offer significant advantages over those currently described in the art. As illustrated in Schemes 1-6, these dyes are designed to prevent aggregation in solution by preventing intramolecular and intermolecular ordered hydrophobic interactions. They also have multiple attachment sites proximal to the dye chromophore for ease of forming dendrimers. The presence of rigid and extended chromophore backbone enhances fluorescence quantum yield and extends the maximum absorption beyond 800 nm. Conjugation of biomolecules to these dyes are readily achievable. They are useful in various biomedical applications including, but not limited to, tomographic imaging of organs; monitoring of organ functions; coronary angiography; fluorescence endoscopy; detection, imaging, and therapy of tumors; laser guided surgery, photoacoustic and sonofluorescence methods; and the like. Specific embodiments to accomplish some of the aforementioned biomedical applications are given below.
In one embodiment of the invention, the dyes of the invention are useful for optical tomographic, endoscopic, photoacoustic and sonofluoresence applications for the detection and treatment of tumors and other abnormalities.
In another aspect of the invention, the dyes of the invention are useful for localized therapy. Typical therapeutic procedures for which the dyes of this invention are useful comprises photodynamic therapy and laser assisted guided surgery (LAGS) for the detection of micrometastases. Also, a combination of imaging and therapy can be accomplished wherein said imaging and therapy consists of absorption, light scattering, photoacoustic, and sonofluoresence techniques.
In yet another aspect of the invention, the dyes of the invention are useful for the detection of the presence of tumors and other abnormalities by monitoring the blood clearance profile of the dyes.
In a further embodiment of the invention, the dyes are useful for laser assisted guided surgery for the detection of micrometastases of tumors upon laparoscopy.
In yet another aspect of the invention, the dye bioconjugates of the dyes of this invention are useful diagnosis of atherosclerotic plaques and blood clots.
The novel dyes of the present invention are prepared according the methods well known in the art and are shown in Schemes 1-5 and their use in the synthesis of bioconjugates is shown in Scheme 6. 
In a preferred embodiment, the dyes according to the present invention have the general Formula 1 wherein A1 and b1 vary from 0 to 3; Q1 is a single bond; R1 to R9 are hydrogens; W1 and X1 may be the same or different and are selected from the group consisting of xe2x80x94C(CH3)2, C((CH2)zzOH)CH3, C((CH2)zzOH)2, C((CH2)zzCO2H)CH3, C((CH2)zzCO2H)2, C((CH2)zzNH2)CH3, C((CH2)zzNH2)2, C((CH2)zzNRppRpz)CH3 and C((CH2)zzNRppRpz)2; Y1 and Z1 may be the same or different and are selected from the group consisting of xe2x80x94(CH2)cxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)dxe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)gxe2x80x94N(R14)xe2x80x94(CH2)hxe2x80x94CO2H, and xe2x80x94(CH2)ixe2x80x94N(R15)xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)jxe2x80x94CH2xe2x80x94CO2H; Rpp and Rpz may be the same or different and are selected from the group consisting of xe2x80x94(CH2)cxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)dxe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)gxe2x80x94N(R14)xe2x80x94(CH2)hxe2x80x94CO2H, and xe2x80x94(CH2)ixe2x80x94N(R15)xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)jxe2x80x94CH2xe2x80x94CO2H; R14 and R15 may be same or different and are selected from the group consisting of -hydrogen, C1-C10 alkyl, C1-C10 aryl, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, xe2x80x94CH2(CH2xe2x80x94Oxe2x80x94CH2)cxe2x80x94CH2xe2x80x94OH, C1-C20 polyhydroxyalkyl, C1-C10 polyhydroxyaryl, xe2x80x94(CH2)dxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)exe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)fNH2, and xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)gxe2x80x94CH2xe2x80x94NH2; c, e, g, h, i, and zz vary from 1 to 5; and d, f and j vary from 1 to 100.
In another preferred embodiment, the dyes according to the present invention have the general Formula 2 wherein a2 and b2 vary from 0 to 3; Q2 is a single bond; R16 to R28 are hydrogens; W2 and X2 may be the same or different and are selected from the group consisting of xe2x80x94C(CH3)2, C((CH2)zzOH)CH3, C((CH2)zzOH)2, C((CH2)zzCO2H)CH3, C((CH2)zzCO2H)2, C((CH2)zzNH2)CH3, C((CH2)zzNH2)2, C((CH2)zzNRppRpz)CH3 and C((CH2)zzNRppRpz)2; Y2 and Z2 may be the same or different and are selected from the group consisting of xe2x80x94(CH2)cxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)dxe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)gxe2x80x94N(R14)xe2x80x94(CH2)hxe2x80x94CO2H, and xe2x80x94(CH2)ixe2x80x94N(R15)xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)jxe2x80x94CH2xe2x80x94CO2H; Rpp and Rpz may be the same or different and are selected from the group consisting of xe2x80x94(CH2)cxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)dxe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)gxe2x80x94N(R14)xe2x80x94(CH2)hxe2x80x94CO2H, and xe2x80x94(CH2)ixe2x80x94N(R15)xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)jxe2x80x94CH2xe2x80x94CO2H; R14 and R15 may be same or different and are selected from the group consisting of -hydrogen, C1-C10 alkyl, C1-C10 aryl, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, xe2x80x94CH2(CH2xe2x80x94Oxe2x80x94CH2)cxe2x80x94CH2xe2x80x94OH, C1-C20 polyhydroxyalkyl, C1-C10 polyhydroxyaryl, xe2x80x94(CH2)dxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)exe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)fNH2, and xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)gxe2x80x94CH2xe2x80x94NH2; c, e, g, h, i, and zz vary from 1 to 5; and d, f and j vary from 1 to 100.
In another preferred embodiment, the dyes according to the present invention have the general Formula 3 wherein a3 and b3 vary from 0 to 3; A1 is a single bond; B1 is selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, and xe2x80x94NR38; C1 is xe2x80x94CH2 or xe2x80x94Cxe2x95x90O; D1 is selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, and xe2x80x94NR38; R29 is a hydrogen, a halogen atom, a saccharide or a hydrophilic peptide; R30 to R37 are hydrogens; R38 is selected from the group consisting of -hydrogen, C1-C10 alkyl, C1-C10 aryl, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, xe2x80x94CH2(CH2xe2x80x94Oxe2x80x94CH2)cxe2x80x94CH2xe2x80x94OH, C1-C20 polyhydroxyalkyl, C1-C10 polyhydroxyaryl, xe2x80x94(CH2)dxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)exe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)fxe2x80x94NH2, and xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)gxe2x80x94CH2xe2x80x94NH2; W3 and X3 may be the same or different and are selected from the group consisting of xe2x80x94C(CH3)2, C((CH2)zzOH)CH3, C((CH2)zzOH)2, C((CH2)zzCO2H)CH3, C((CH2)zzCO2H)2, C((CH2)zzNH2)CH3, C((CH2)zzNH2)2, C((CH2)zzNRppRpz)CH3 and C((CH2)zzNRppRpz)2; Y3 and Z3 may be the same or different and are selected from the group consisting of xe2x80x94(CH2)cxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)dxe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)gxe2x80x94N(R14)xe2x80x94(CH2)hxe2x80x94CO2H, and xe2x80x94(CH2)ixe2x80x94N(R15)xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)jxe2x80x94CH2xe2x80x94CO2H; Rpp and Rpz may be the same or different and are selected from the group consisting of xe2x80x94(CH2)cxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)dxe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)gxe2x80x94N(R14)xe2x80x94(CH2)hxe2x80x94CO2H, and xe2x80x94(CH2)ixe2x80x94N(R15)xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)jxe2x80x94CH2xe2x80x94CO2H; R14 and R15 may be same or different are selected from the group consisting of -hydrogen, C1-C10 alkyl, C1-C10 aryl, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, xe2x80x94CH2(CH2xe2x80x94Oxe2x80x94CH2)cxe2x80x94CH2xe2x80x94OH, C1-C20 polyhydroxyalkyl, C1-C10 polyhydroxyaryl, xe2x80x94(CH2)dxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)exe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)fNH2, and xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)gxe2x80x94CH2xe2x80x94NH2; c, e, g, h, i, and zz vary from 1 to 5; and d, f and j vary from 1 to 100.
In another preferred embodiment, the dyes according to the present invention have the general Formula 4 wherein a4 and b4 vary from 0 to 3; A2 is a single or double bond; B2 is selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, or xe2x80x94NR38; C2 is xe2x80x94CH2 or xe2x80x94Cxe2x95x90O; D2 is selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94 and xe2x80x94NR38; R38 is selected from the group consisting of -hydrogen, C1-C10 alkyl, C1-C10 aryl, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, xe2x80x94CH2(CH2xe2x80x94Oxe2x80x94CH2)cxe2x80x94CH2xe2x80x94OH, C1-C20 polyhydroxyalkyl, is C1-C10 polyhydroxyaryl, xe2x80x94(CH2)dxe2x80x94CO2H, CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)exe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)fxe2x80x94NH2, and xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)gxe2x80x94CH2xe2x80x94NH2; R45 is a hydrogen, a halogen atom, a saccharide or a hydrophilic peptide; R46 to R57 are hydrogens; W4 and X4 may be the same or different and are selected from the group consisting of xe2x80x94C(CH3)2, C((CH2)zzOH)CH3, C((CH2)zzOH)2, ((CH2)zzCO2H)CH3, C((CH2)zzCO2H)2, C((CH2)zzNH2)CH3, C((CH2)zzNH2)2, C((CH2)zzNRppRpz)CH3 and C((CH2)zzNRppRpz)2; Y4 and Z4 may be the same or different and are selected from the group consisting of xe2x80x94(CH2)cxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)dxe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)gxe2x80x94N(R14)xe2x80x94(CH2)hxe2x80x94CO2H, and xe2x80x94(CH2)ixe2x80x94N(R15)xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)jxe2x80x94CH2xe2x80x94CO2H; Rpp and Rpz are the same or different and are selected from xe2x80x94(CH2)cxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)dxe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)gxe2x80x94N(R14)xe2x80x94(CH2)hxe2x80x94CO2H, and xe2x80x94(CH2)ixe2x80x94N(R15)xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)jxe2x80x94CH2xe2x80x94CO2H; R14 and R15 may be the same or different and are selected from the group consisting of -hydrogen, C1-C10 alkyl, C1-C10 aryl, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, xe2x80x94CH2(CH2xe2x80x94Oxe2x80x94CH2)cxe2x80x94CH2xe2x80x94OH, C1-C20 polyhydroxyalkyl, C1-C10 polyhydroxyaryl, xe2x80x94(CH2)dxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)exe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)fxe2x80x94NH2, and xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)gxe2x80x94CH2xe2x80x94NH2; c, e, g, h, i, and zz vary from 1 to 5; and d, f and j vary from 1 to 100.
In another preferred embodiment, the dyes according to the present invention have the general Formula 5 wherein a5 varies from 0 to 3; A3 is a single or double bond; B3 is selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94 and xe2x80x94NR38; C3 is xe2x80x94CH2 or xe2x80x94Cxe2x95x90O; D3 is selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94 and xe2x80x94NR38; R38 is selected from the group consisting of -hydrogen, C1-C10 alkyl, C1-C10 aryl, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, xe2x80x94CH2(CH2xe2x80x94Oxe2x80x94CH2)cxe2x80x94CH2xe2x80x94OH, C1-C20 polyhydroxyalkyl, C1-C10 polyhydroxyaryl, xe2x80x94(CH2)dxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)exe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)fxe2x80x94NH2, and xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)gxe2x80x94CH2xe2x80x94NH2; R58 is a hydrogen, a halogen atom, a saccharide or a hydrophilic peptide; R59 to R66 are hydrogens; W5 and X5 may be the same or different and are selected from the group consisting of xe2x80x94C(CH3)2, C((CH2)zzOH)CH3, C((CH2)zzOH)2, C((CH2)zzCO2H)CH3, C((CH2)zzCO2H)2, C((CH2)zzNH2)CH3, C((CH2)zzNH2)2, C((CH2)zzNRppRpz)CH3 and C((CH2)zzNRppRpz)2; Y5 and Z5 may be the same or different and are selected from the group consisting of xe2x80x94(CH2)cxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)dxe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)gxe2x80x94N(R14)xe2x80x94(CH2)hxe2x80x94CO2H, and xe2x80x94(CH2)ixe2x80x94N(R15)xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)jxe2x80x94CH2xe2x80x94CO2H; Rpp and Rpz are the same or different and are selected from xe2x80x94(CH2)cxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)dxe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)gxe2x80x94N(R14)xe2x80x94(CH2)hxe2x80x94CO2H, and xe2x80x94(CH2)ixe2x80x94N(R15)xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)jxe2x80x94CH2xe2x80x94CO2H; R14 and R15 may be the same or different and are selected from the group consisting of -hydrogen, C1-C10 alkyl, C1-C10 aryl, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, xe2x80x94CH2(CH2xe2x80x94Oxe2x80x94CH2)cxe2x80x94CH2xe2x80x94OH, C1-C20 polyhydroxyalkyl, C1-C10 polyhydroxyaryl, xe2x80x94(CH2)dxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)exe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)fNH2, and xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)gxe2x80x94CH2xe2x80x94NH2; c, e, g, h, i, and zz vary from 1 to 5; and d, f and j vary from 1 to 100.
In another preferred embodiment, the dyes according to the present invention have the general Formula 6 wherein a6 varies from 0 to 3; A4 is a single or double bond; B4 is selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94 and xe2x80x94NR38; C4 is xe2x80x94CH2 or xe2x80x94Cxe2x95x90O; D4 is selected from the group consisting of xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94 and xe2x80x94NR38; R38 is selected from the group consisting of -hydrogen, C1-C10 alkyl, C1-C10 aryl, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, xe2x80x94CH2(CH2xe2x80x94Oxe2x80x94CH2)cxe2x80x94CH2xe2x80x94OH, C1-C20 polyhydroxyalkyl, C1-C10 polyhydroxyaryl, xe2x80x94(CH2)dxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)exe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)fNH2, and xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)gxe2x80x94CH2xe2x80x94NH2; R67 is a hydrogen, a halogen atom, a saccharide or a hydrophilic peptide; R68 to R79 are hydrogens; W6 and X6 may be the same of different and are selected from the group consisting of xe2x80x94C(CH3)2, C((CH2)zzOH)CH3, C((CH2)zzOH)2, C((CH2)zzCO2H)CH3 , C((CH2) CO2H)2, C((CH2)zzNH2)CH3, C((CH2)zzNH2)2, C((CH2)zzNRppRpz)CH3 and C((CH2)zzNRppRpz)2; Y6 and Z6 may be the same or different and are selected from the group consisting of xe2x80x94(CH2)cxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)dxe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)gxe2x80x94N(R14)xe2x80x94(CH2)hxe2x80x94CO2H and xe2x80x94(CH2)ixe2x80x94N(R15)xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)jxe2x80x94CH2xe2x80x94CO2H; Rpp and Rpz are the same or different and are selected from the group consisting of xe2x80x94(CH2)cxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)dxe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)gxe2x80x94N(R14)xe2x80x94(CH2)hxe2x80x94CO2H and xe2x80x94(CH2)ixe2x80x94N(R15)xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2jxe2x80x94CH2xe2x80x94CO2H; R14 and R15 may be the same different and are selected from the group consisting of -hydrogen, C1-C10 alkyl, C1-C10 aryl, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, xe2x80x94CH2(CH2xe2x80x94Oxe2x80x94CH2)cxe2x80x94CH2xe2x80x94OH, C1-C20 polyhydroxyalkyl, C1-C10 polyhydroxyaryl, xe2x80x94(CH2)dxe2x80x94CO2H, xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)exe2x80x94CH2xe2x80x94CO2H, xe2x80x94(CH2)fNH2, and xe2x80x94CH2xe2x80x94(CH2xe2x80x94Oxe2x80x94CH2)gxe2x80x94CH2xe2x80x94NH2; c, e, g, h, i, and zz vary from 1 to 5; and d, f and j vary from 1 to 100.
The compositions of the invention can be formulated into diagnostic compositions for enteral or parenteral administration. These compositions contain an effective amount of the dye along with conventional pharmaceutical carriers and excipients appropriate for the type of administration contemplated. For example, parenteral formulations advantageously contain a sterile aqueous solution or suspension of dye according to this invention. Parenteral compositions may be injected directly or mixed with a large volume parenteral composition for systemic administration. Such solutions also may contain pharmaceutically acceptable buffers and, optionally, electrolytes such as sodium chloride.
Formulations for enteral administration may vary widely, as is well known in the art. In general, such formulations are liquids which include an effective amount of the dye in aqueous solution or suspension. Such enteral compositions may optionally include buffers, surfactants, thixotropic agents, and the like. Compositions for oral administration may also contain flavoring agents and other ingredients for enhancing their organoleptic qualities.
The diagnostic compositions are administered in doses effective to achieve the desired enhancement. Such doses may vary widely, depending upon the particular dye employed, the organs or tissues which are the subject of the imaging procedure, the imaging equipment being used, and the like.
The diagnostic compositions of the invention are used in the conventional manner. The compositions may be administered to a patient, typically a warm-blooded animal, either systemically or locally to the organ or tissue to be imaged, and the patient then subjected to the imaging procedure.